Apical localization of the coxsackie-adenovirus receptor by glycosyl-phosphatidylinositol modification is sufficient for adenovirus-mediated gene transfer through the apical surface of human airway epithelia

R. W. Walters, W. Van't Hof, S. M.P. Yi, M. K. Schroth, J. Zabner, Ronald Crystal, M. J. Welsh

Research output: Contribution to journalArticle

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Abstract

In well-differentiated human airway epithelia, the coxsackie B and adenovirus type 2 and 5 receptor (CAR) resides primarily on the basolateral membrane. This location may explain the observation that gene transfer is inefficient when adenovirus vectors are applied to the apical surface. To further test this hypothesis and to investigate requirements and barriers to apical gene transfer to differentiated human airway epithelia, we expressed CAR in which the transmembrane and cytoplasmic tail were replaced by a glycosyl-phosphatidylinositol (GPI) anchor (GPI-CAR). As controls, we expressed wild-type CAR and CAR lacking the cytoplasmic domain (Tailless-CAR). All three constructs enhanced gene transfer with similar efficiencies in fibroblasts. In airway epithelia, GPI-CAR localized specifically to the apical membrane, where it bound adenovirus and enhanced gene transfer to levels obtained when vector was applied to the basolateral membrane. Moreover, GPI-CAR facilitated gene transfer of the cystic fibrosis transmembrane conductance regulator to cystic fibrosis airway epithelia, correcting the Cl- transport defect. In contrast, when we expressed wild-type CAR it localized to the basolateral membrane and failed to increase apical gene transfer. Only a small amount of Tailless-CAR resided in the apical membrane, and the effects on apical virus binding and gene transfer were minimal. These data indicate that binding of adenovirus to an apical membrane receptor is sufficient to mediate effective gene transfer to human airway epithelia and that the cytoplasmic domain of CAR is not required for this process. The results suggest that targeting apical receptors in differentiated airway epithelia may be sufficient for gene transfer in the genetic disease cystic fibrosis.

Original languageEnglish
Pages (from-to)7703-7711
Number of pages9
JournalJournal of Virology
Volume75
Issue number16
DOIs
Publication statusPublished - 22 Aug 2001
Externally publishedYes

Fingerprint

Coxsackie and Adenovirus Receptor-Like Membrane Protein
Glycosylphosphatidylinositols
phosphatidylinositols
Adenoviridae
gene transfer
epithelium
Epithelium
receptors
Genes
Membranes
cystic fibrosis
Cystic Fibrosis
Virus Attachment
Cystic Fibrosis Transmembrane Conductance Regulator
Inborn Genetic Diseases
genetic disorders
fibroblasts
Tail
tail
Fibroblasts

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Apical localization of the coxsackie-adenovirus receptor by glycosyl-phosphatidylinositol modification is sufficient for adenovirus-mediated gene transfer through the apical surface of human airway epithelia. / Walters, R. W.; Van't Hof, W.; Yi, S. M.P.; Schroth, M. K.; Zabner, J.; Crystal, Ronald; Welsh, M. J.

In: Journal of Virology, Vol. 75, No. 16, 22.08.2001, p. 7703-7711.

Research output: Contribution to journalArticle

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